VIP® Gold for Listeria
AOAC Official Method 997.03
AOAC Performance Tested Method 060801
GENERAL DESCRIPTION
VIP® Gold for Listeria is a single-step visual immunoassay for the
detection of Listeria in food and environmental samples. Each
device contains a proprietary reagent system, which forms a
visually apparent antigen-antibody-chromogen complex if Listeria
is present. The test is intended for use by laboratory personnel
with appropriate microbiology training.
Part No: 60037-40 (40 tests)
KIT COMPONENTS
Each VIP Gold for Listeria kit contains the following:
VIP Gold for Listeria test devices
EQUIPMENT / MATERIALS REQUIRED
Other necessary materials not provided include:
Media per Appendix A
Autoclave
Vortex mixer
Analytical balance, tolerance ± 0.2 g
Stomacher / Masticator machine
Stomacher-type bags with filter or equivalent
Incubator capable of maintaining 29-31°C
Water bath capable of maintaining 95-105°C or
equivalent (e.g. autoclave with flowing steam, dry heater)
Micropipette(s) capable of delivering 0.1 mL and 1.0 mL
SAMPLE PREPARATION
A. Test Portion Preparation & Enrichment
Two Step Enrichment Protocol (AOAC OMA 997.03)
for meat, poultry, seafood, vegetables, dairy products, eggs,
pasta, animal meal, nuts and environmental samples.
(a) Food samples Add 25 g test portion to 225 mL of
modified Fraser Broth with lithium chloride (mFB+LiCl)
(Appendix A). Stomach / masticate for 2 minutes and
incubate 28 h (26-30 h) at 30°C (29-31°C).
Environmental samples Add 60 mL of mFB+LiCl to
sample bag containing environmental sponge sample. Ensure
that the sponge is oriented horizontally in the sample bag. If
using a swab, add environmental swab sample to 10 mL of
mFB+LiCl. Mix well and incubate 28 h (26-30 h) at 30°C
(29-31°C).
(b) Transfer 1 mL incubated mFB+LiCl to 9 mL Buffered
Listeria Enrichment Broth (BLEB) (Appendix A). Mix
thoroughly with vortex mixer. Incubate 24 h (22-26 h) at
30°C (29-31°C).
Single Step Enrichment Protocol (AOAC PTM 060801)
for meat, poultry, seafood, vegetables, dairy products, and
environmental samples.
(a) Food samples Add 25 g test portion to 225 mL of Demi-
Fraser Broth (DFB) (Appendix A). Stomach / masticate for
2 minutes and incubate 48 h (46-54 h) at 30°C (29-31°C).
(b) Environmental samples Add 100 mL of DFB to sample
bag containing environmental sponge sample. Ensure that
the sponge is oriented horizontally in the sample bag. If
using a swab, add environmental swab sample to 10 mL of
DFB. Mix well and incubate 48 h (46-54 h) at 30°C (29-
31°C).
Note: Retain original BLEB or DFB tubes under
refrigeration (2-8°C). Use for confirmation of presumptive
positive results.
B. Sample Inactivation
(a) Vortex mix incubated sample. Transfer 1.0 mL incubated
BLEB or DFB to a clean test tube.
(b) Inactivate microorganisms at 100°C (95-105°C) for 5 min
in a water bath, autoclave or dry heater.
(c) Cool tubes to 25-37°C before testing. Tubes that have
been inactivated can be stored for up to 4 days at 2-8°C
prior to testing.
TEST PROCEDURE
(a) Open the sealed foil pouch containing the VIP Gold
devices and remove the sheet of test devices. Break away
the necessary number of devices, one device for each
test portion.
(b) VIP Gold units may not be reused. Reseal unused VIP
Gold units in pouch containing desiccant. Store at room
temperature (15-25°C).
(c) Enriched broths should be equilibrated to 25-37°C prior
to running the test.
(d) Vortex mix contents. Transfer 0.1 mL of inactivated
BLEB or DFB to sample addition well. Avoid transferring
particulate matter to the device.
(e) Incubate device at room temperature (15-25°C) for 10
min.
RESULTS
Note: Examine test unit at 10 min. Beyond this time faint
lines may develop because of non-specific color development
and should be disregarded.
(a) Examine VIP Gold unit for the presence of distinct
detection lines in both test sample and test verification
zone. Lines should be dark when contrasted with white
background and should extend across the zone. Intensity
of test sample and test verification lines may differ.
Absence of a test verification line indicates an invalid test
result. Contact BioControl Technical Services at (800)
245-0113.
(b) Test sample is considered positive when lines are
present in the test sample zone and in the test
verification zone.
Sample
addition well
Sample Verification
zone zone
(c) Test sample is considered negative when a line is
present in the test verification zone and no line is seen in
the test sample zone.
Sample
addition well
Sample Verification
zone zone
(d) Positive and negative control cultures should be run to
familiarize the analyst with the results interpretation.
CONFIRMATION
Presumptive positive samples should be confirmed from the
retained BLEB or DFB tubes via either:
US FDA. 2003. Bacteriological Analytical Manual online, chapter
10. http://www.cfsan.fda.gov/~ebam/bam-toc.html
USDA-FSIS. 2008. Microbiology Laboratory Guidebook, chapter 8.
http://www.fsis.usda.gov/science/microbiological_lab_guidebook/
index.asp
STORAGE CONDITIONS
APPENDIX A
Store VIP Gold for Listeria kit components at room
temperature (15-25°C). Do not refrigerate. Modified Fraser Broth with Lithium Chloride
(mFB+LiCl)
Reseal unused VIP Gold units in pouch with desiccant pack.
The moisture indicator line on the desiccant pack must be Suspend 55 g of Fraser Broth Base (FB) in 1 L deionized
blue. water. Mix thoroughly until completely dissolved. Add 4 g
lithium chloride (LiCl) and stir until completely dissolved.
Kit expiration is provided on the product pouch label.
Autoclave at 121ºC for 15 min. Do not overheat. Do not add
PRECAUTIONS ferric ammonium citrate additive to broth. Do not use broth
VIP Gold for Listeria must be used as described herein. This if precipitate forms. Alternately, prepare a 45% (w/v) LiCl
product is not intended for human or veterinary use. Do not solution by dissolving 45 g LiCl in enough deionized water to
use test kit beyond expiration date indicated on the label. Do
make a final volume of 100 mL. Filter sterilize through a 0.2
no mix reagents from different Assurance kit lots.
µm filter. Add 2 mL sterile LiCl solution to 225 mL of
Avoid contact with eyes, skin, and clothing. Avoid swallowing sterilized FB. If using commercially prepared 8 M LiCl
or taking product internally.
solution, add 2.65 mL per 225 mL sterilized FB.
Decontaminate materials by autoclave, bleach, etc., in
accordance with good laboratory practices and in accordance Buffered Listeria Enrichment Broth (BLEB)
with local, state and federal regulations. Waste may be Suspend 36.1 g Listeria Enrichment Broth in 1 L deionized
contaminated with Listeria which is potentially hazardous to water. Add 8.5 g 3-3-(N-Morpholino) propanesulfonic acid
human health. All biohazard waste should be disposed of (MOPS) free acid and 13.7 g MOPS sodium salt. Mix
appropriately.
thoroughly. Autoclave at 121°C for 15 min.
Pregnant women, elderly, and potentially
immunocompromised individuals must be prohibited from Demi-Fraser Broth (DFB)
laboratory rooms or areas where L. monocytogenes Suspend 55 g of Demi-Fraser Broth Base in 1 L deionized
enrichment, isolation, and identification procedures are in water. Mix thoroughly until completely dissolved. Autoclave
progress. Although a properly sanitized laboratory area at 121ºC for 15 min. Do not add ferric ammonium citrate
should not harbor Listeria, supervisors should use their own
discretion in allowing these high-risk individuals into these additive to broth.
areas.
PRODUCT WARRANTY
BioControl Systems, Inc. (BCS) warrants this product to be free from defects
in materials and workmanship, when stored under label conditions and used
as intended until the expiration date stated on the package. BCS agrees
during the applicable warranty period to replace all defective products after
return to BCS. BCS shall not have obligation under this Limited Warranty to
make replacements which result, in whole or in part, from negligence of the
Buyer, or from improper use of the products, or use of the product in a
manner for which it was not indicated. Buyer shall notify BCS of any
products which it believes to be defective during the Warranty period. At
BCS option, such products shall be returned to BCS, transportation and
insurance prepaid. BCS shall replace any such product found to be defective,
at no charge. Should BCS examination not disclose any defect covered by the
foregoing warranty, BCS shall so advise Buyers and dispose of the product in
accordance with Buyer’s instructions.

BioControl Systems, Inc. 2008 BioControl Systems, Inc., certifies that VIP
®
Gold for Listeria meets all requirements specified in AOAC Official Method
997.03 Listeria monocytogenes and related Listeria spp. in Selected Foods,
Visual Immunoprecipitate Assay. BioControl and VIP are registered
trademarks of BioControl Systems, Inc. US patent #5,658,747, European
patent #0,759,172. Canada patent #2,190,109. Australia patent #680362.
Japan patent #2894841. Mexico Patent #230973. Other US and international
patents are pending. Printed in USA. All rights reserved.

55167.R000.042009
Part No: 60037-40